Electroporation and GFP-labelled transplantation of testicular cells in Nile tilapia

  • Epro Barades Sekolah Pascasarjana, Program Studi Ilmu Akuakultur, Institut Pertanian Bogor, Kampus IPB Dramaga Bogor 16680
  • , Alimuddin Departemen Budidaya Perairan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor Kampus IPB Dramaga Bogor 16680
  • Agus Oman Sudrajat Departemen Budidaya Perairan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor Kampus IPB Dramaga Bogor 16680
DOI: https://doi.org/10.19027/jai.12.186-192

Abstract

ABSTRACT

 

Transplantation technology can be applied to generate fish surrogate broodstock. A donor germinal cells that was used in transplantation are labeled to distinguish it with endogenous cells. Donor cells are generally derived from transgenic fish carrying a marker or cells labeled by PKH-26. This study was performed to obtain an alternative method of cell labelling using electroporation. Testicular cells were taken from 4-months old Nile tilapia as a model. Electroporation was performed with testicular cell density of 104 cells/µL, pJfKer-GFP concentration of 50 ng/µL, and a pulse length of 20 ms at 0, 100, 200, and 300 volts. At amount of 5x103 cells/0.5µL electroporated testicular cells were then injected into the intraperitoneal cavity of 3-day-old Nile tilapia larva. The results showed that survival of the electroporated cells of 100 and 200 volt-treatments was similar (P>0.05), and higher than 300 volt (P<0.05). Number of fluorescent cells was not significantly different among treatments. The highest cell colonization in transplanted fish was obtained in 200-volt treatment (66.67%). As conclusion, 200-volt electroporation with was a suitable tool to label testicular cells for transplantation.

 

Keyword: electroporation, GFP, label, Nile tilapia, transplantation

 

 

ABSTRAK

 

Teknologi transplantasi merupakan suatu teknologi yang dapat menghasilkan induk pengganti. Sel donor berupa sel germinal yang akan digunakan dalam transplantasi diberi label agar dapat dibedakan dengan sel resipien. Umumnya sel donor diperoleh dari ikan transgenik yang membawa marka atau diwarnai dengan PKH-26. Penelitian ini bertujuan untuk mencari metode alternatif dalam pemberian label pada sel dengan elektroporasi. Sel testikular diperoleh dari ikan nila berumur empat bulan sebagai model. Elektroporasi dilakukan dengan kepadatan sel 104 sel/µL, konsentrasi pJfKer-GFP 50 ng/µL, dan panjang kejut 20 ms pada 0, 100, 200, dan 300 volt. Sebanyak 5x103 sel dalam 0,5 µL larutan hasil elektroporasi disuntikkan ke dalam rongga intraperitoneal larva berumur tiga hari setelah menetas. Hasil penelitian menunjukkan bahwa kelangsungan hidup sel pada kejut 100 dan 200 volt tidak signifikan (P<0,05), akan tetapi lebih tinggi bila dibandingkan dengan perlakuan 300 volt (P<0,05). Jumlah sel yang berpendar tidak berbeda antara perlakuan. Persentase kolonisasi sel pada ikan hasil transplan tertinggi pada perlakuan 200 volt (66,67%). Kesimpulan yang diperoleh dari hasil penelitian adalah perlakuan elektroporasi dengan kejut listrik 200 volt dapat digunakan untuk memberi label pada sel testikular yang akan ditransplantasikan.

 

Kata kunci : elektroporasi, GFP, label, ikan nila, transplantasi

 

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Published
2015-05-12
How to Cite
[1]
BaradesE., Alimuddin,. and SudrajatA.O. 2015. Electroporation and GFP-labelled transplantation of testicular cells in Nile tilapia. Jurnal Akuakultur Indonesia. 12, 2 (May 2015), 186-192. DOI:https://doi.org/10.19027/jai.12.186-192.
Issue
Vol. 12 No. 2 (2013): Jurnal Akuakultur Indonesia
Section
Articles

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